• 제목/요약/키워드: HPLC pattern analysis

검색결과 95건 처리시간 0.025초

한국산 무당개구리 피부에 존재하는 Bombesin 유사면역 반응물질의 순수정제 및 생물학적 활성 (Purification and Biological Activities of Bombesin Like Immunoreactivity from Skin of the Frog, Bombina orientalis in Korea)

  • 권혁일;김일;박형진
    • The Korean Journal of Physiology
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    • 제24권2호
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    • pp.363-375
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    • 1990
  • The present investigation was performed to purify bombesin-like immunoreactivity (BBS-LI) from the skin of frogs, B. orientalis inhabiting Korea. For extraction of BBS-LI, the fresh skin of 360 g from frogs was immersed in 1,800 ml of 100% methanol and then kept at $4^{\circ}C$ for 5 days. BBS-LI was partially purified by liquid chromatography using an alkaline alumina column followed by a Sephadex G-10 column. BBS-LI was further purified by using sequential HPLC of reversed phase C18 preparation, gel permeation, SP-ion exchange and reversed phase C18 analysis. BBS-LI in fractions of each step was monitored by radioimmunoassay for which bombesin antiserum with a titer of 1 : 188,800 was raised in a guinea pig. Eventually, two different BBS-LI were successfully purified and each BBS-LI showed the following character. 1) BBS-LI was well separated into two peaks in SP-ion exchange HPLC. One (BBS-LI-K1) bound to the column while the other (BBS-LI-K2) did not. 2) BBS-LI-K1, 73.8% of total BBS-LI, was not differentiated from synthetic bombesin in reversed phase C18 analytical and gel permeation HPLC. 3) BBS-LI-K2, 26.2% of total BBS-LI, eluted later than synthetic bombesin in reversed phase C18 analytical HPLC, but it eluted with a retention time identical to that of synthetic bombesin in gel permeation HPLC. 4) The two forms of BBS-LI and synthetic bombesin identically stimulated gastrin release and pancreatic exocrine secretion including volume, protein output and amylase output in anesthetized rats. It is concluded from the above results that the skin of B. orientalis contains two different forms of BBS-LI which are very identical to bombesin immunologically and biologically. In comparison with synthetic bombesin containing 14 amino acid residues, the major form shows quite similar pattern in all HPLC used in the present study, but the minor form exhibits quite different pattern in SP-ion exchange and reversed phase C18 analytical HPLG.

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Simultaneous Determination and Recognition Analysis of Coumarins in Angelica decursiva and Peucedanum praeruptorum by HPLC-DAD

  • Kim, Hye Mi;Jeong, Su Yang;Kim, Sun Min;Lee, Kyu Ha;Kim, Jong Hwan;Seong, Rack Seon
    • Natural Product Sciences
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    • 제22권3호
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    • pp.162-167
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    • 2016
  • Peucedani Radix is the root of Angelica decursiva Franchet et Savatier (=Peucedanum decursivum Maximowicz) or Peucedanum praeruptorum Dunn in several Asian countries. The coumarins contained in Peucedani Radix were quantitatively analyzed using HPLC-DAD to develop a simultaneous determination for the quality control of A. decursiva and P. praeruptorum. For quantitative analysis, four major coumarins contained in these medicinal plants were assessed. Nodakenin (1), nodakenetin (2), praeruptorin A (3), and praeruptorin B (4) were separated with a Phenomenex Luna C18 column ($5{\mu}m$, $4.6{\times}250mm$) under the gradient conditions using distilled water with 0.1% phosphoric acid and acetonitrile with 0.1% phosphoric acid as the mobile phase, at a flow rate of 1.0 ml/min and a detection wavelength of 330 nm. This method was fully validated for linearity, accuracy, precision, recovery, and limit of detection and quantification. As a result, A. decursiva and P. praeruptorum were clearly classified by the quantification of four major coumarins in extracts. Also, the pattern recognition analysis based on HPLC indicates that all of the samples were largely clustered into two groups. Therefore, it is possible to distinguish between A. decursiva and P. praeruptorum and contribute to quality control.

4종류 목향류(木香類)의 외부(外部) 및 내부형태(內部形態)와 이화학패턴연구 (A Study on external and internal morphology and pattern analysis in 4 kinds of Mok-Hyaeng Radix)

  • 김홍준;강경식;최고야;김호경;정승일;주영승
    • 한국한의학연구원논문집
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    • 제12권3호통권18호
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    • pp.117-130
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    • 2006
  • The purpose of this study was to introduce the differential standard of Mok-Hyaeng Radix species. We established the classificatory standard according to the external and internal morphology and the pattern in 4 kinds of Mok-Hyaeng Radix. The results follow: 1. On the external morphology, Radix of Vladimiria souliei had a sticky material in the head of roots and it was yellowish white in the cut surface. It attached to teeth during chewing. Radix of Aucklandia lappa (=Saussurea lappa) was dark grayish brown and was grayish brown in the cut surface. It had special odor and was not attache to teeth chewing it. On the other hand, Radix of Inula helenium was dark brown. In the cut surface, cortical layer was different from wood layer. Radix of Aristolochia contorta was yellowish brown. In the cut surface, it was not smooth and cortical layer was different from wood layer. 2. On the internal morphology, Radix of Vladimiria souliei had many fascicular fibers and scattered large oil chambers in the bast ray, xylem ray, and parenchyma. On Radix of Aucklandia lappa (=Saussurea lappa), it was difficult to find out the trace when the cork layer was exfoliated. The cambium formed circle and the large oil chambers were scattered in the phloem and xylem. On the other hand, external cortical layer of Radix of Inula helenium was detached and cambium was formed to indistinct circle and large oil chambers were scattered in the phloem and xylem. On Radix of Arustolochia contorta, the large oil chambers were scattered in the surface later, bast part was relatively broad and cambium formed circle. 3. On the TLC pattern, Radix of Aristolochia contorta, Inula helenium, Aucklandia lappa, and Vladimiria souliei were chromogenic to violet and purple according to sulfuric acid colour pattern, especially faint on Aristolochia contorta. 4. On the pattern analysis of HPLC, we compared the content and the pattern of constunolide and dehydrocostus lactone, Radix of Aucklandia lappa was similar to Radix of Vladimiria souliei and Radix of Aristolochia contorta was similar to Radix of Inula helenium. On the total HPLC component pattern, constunolide and dehydrocostus lactone were similar in all four types, but it showed the Rf 5-6 peak was only in Radix of Aristolochia contorta, not in Radix of Inula helenium, Aucklandia lappa, and Vladimiria souliei. It is considered the results of this study will be furnished the basis to succeeding studies and it is needed to extensive comparative study for the same genus-degree of relatedness.

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자소엽(紫蘇葉)의 한약재 표준화 연구 (Analysis of Perilla Frutescens Using Liquid Chromatogram Pattern)

  • 김동우;황귀서
    • 대한예방한의학회지
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    • 제10권2호
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    • pp.131-145
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    • 2006
  • Perilla frutescens is known as the herb helps digestion, tonifies stomachache, and decreases fever in oriental medicine. And it is reported it possess the anti-pyretic effect, anti-inflammatory effect, anti-allergy effect, anti-tumor effect etc. The components isolated from this herb consist of perilla aldehyde, d-limonene, ${\alpha}-pinene$, cyanin, linoleic acid, palmitic acid, menthol, rosmarinic acid and luteolin etc. But there is no effective tools to determine the quality of this herb. In this study, we aimed to analyze the changes of liquid chromatogram pattern, one of major standardization method, to determine the quality of Perilla frutescens.

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국내산 아까시나무와 밤나무 유래 propolis의 영양성분 및 페놀성 화합물 분석 (Analysis of Nutritional Composition and Phenolic Compound in Propolis Collected from Falseacacia and Chestnut Tree in Korea)

  • 송효남;길복임
    • 한국식품과학회지
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    • 제34권4호
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    • pp.546-551
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    • 2002
  • 국내에서 채집되는 propolis에 대한 기초자료를 얻고자 아까시나무와 밤나무를 밀원 식물로 하여 채집된 propolis와 그의 70% ethanol 추출물(EEP)에 대한 영양성분 및 페놀성 화합물을 분석하였다. 일반성분은 밀원식물에 따른 큰 차이 없이 비슷한 분포를 보였고, EEP의 경우 propolis 분말에서보다 대부분의 성분들이 상당량 감소되었다. 주요 무기질 5종은 일반성분과는 달리 채집지역과 수종에 따른 차이를 보였고, 대체로 아까시나무의 propolis가 무기물 함량이 많았다. 아까시나무의 propolis에는 칼슘함량이 가장 많았고, 밤나무 propolis는 칼륨이 가장 많이 함유되어 있었다. 반면에 EEP에서는 두 시료 모두 칼륨의 함량이 가장 많았다. 아미노산은 총 16종이 검출되었고, aparagine, cysteine, glutamine 및 tryptophan은 검출되지 않았다. 특히 밤나무 propolis에서는 methionine도 검출되지 않았다. Aspartic acid가 가장 많았고, methionine의 함유량은 극히 적었으며, 아까시나무 propolis가 아미노산 함유량이 전반적으로 더 높았다. 추출수율이 비교적 높게 나타난 EEP의 total polyphenol 함량은 $13.9{\sim}23.7%$였고, propolis의 phenolic fraction중 대부분을 차지하는 성분인 flavonoids 함량은 $8.6{\sim}10.8%$로 나타나 다른 식품재료에 비하여 매우 높은 수준이었다. 고성능 박막크로마토그래피로 페놀성 화합물을 정성분석한 결과 시료종류에 상관없이 거의 동일한 pattern의 chromatogram을 나타내었다. Polyphenol 화합물의 HPLC 분석에서는 약 16개의 peak가 검출되었고, 이중 p-hydroxy benzoic acid, caffeic acid, ferulic acid, benzoic acid, cinnamic acid 및 chrysin의 6종을 동정하였다. 시료간의 chromatographic pattern은 동일하여 propolis를 구성하는 페놀성 화합물의 종류는 큰 차이가 없는 것으로 판단되었다.

Quality Monitoring of Distributed Schizandra chinensis (Turcz.) Baill in Korea

  • Choo, Byung-Kil;Kim, Hong-Jun;Lee, A-Yeong;Chun, Jin-Mi;Lee, Hye-Won;Woo, Myeong-Ji;Kim, Sun-Hee;Kim, Ho-Kyoung
    • 대한한의학회지
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    • 제27권4호
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    • pp.150-155
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    • 2006
  • Object : This study investigated the quality inspection of Schizandra chinensis (Turcz.) Bail distributed in Korea. Methods: To evaluate the quality of these herbal medicines, we carried out TLC pattern analysis, purity, loss on drying, ash, acid-insoluble ash, essential oil contents, dilute ethanol-soluble, water-soluble, ether-soluble extracts and HPLC analysis. Results : As a result, TCL pattern analysis of schizandrin and gomisin A showed Rf=0.64 and 0.74, respectively. To measured content of schizandrin and gomisin A, we quantitatively analyzed using HPLC. The average contents of schizandrin and gomisin A were detected to be 0.60 (${\pm}$0.02)% and 0.12 (${\pm}$0.004)%, respectively. Discussion : As a result of this study, we suggest that the minimum content of schizandrin and gomisin A should be 0.5% and 0.1%, respectively. We could suggest that the minimum of essential oil content should be 0.6 ml.

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Discrimination of Ginseng Habitat by Using Instrumental Analysis Techniques

  • Sohn H. J.;Lee S. K.;Cho B. G.;Kim S. J.;Lee N. Y.;Choi D. S.;Jeong M. S.;Bae H. R.;Yang J. W.
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 2002년도 학술대회지
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    • pp.238-252
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    • 2002
  • In order to screen out indicators for the discrimination of ginseng habitat, some physical and chemical characteristics of Korean red ginsengs (94 kinds) and Chinese red ginsengs (50 kinds) were analyzed by using a rheometer, an electronic nose system, a combined technique of solid phase micro-extraction (SPME) and gas chromatograph equipped with an electron capture detector (GC/ECD), an X-ray fluorescence spectrometer (XRF), an inductively coupled plasma mass spectrometer (ICP/MS), a near infrared spectrometer (NIRs) and high performance liquid chromatography equipped with evaporative light scattering detector (HPLC/ELSD). The results are summarized as follows: (i) The rhizome strengths of Korean red ginsengs were significantly higher than those of Chinese red ginsengs. (ii) The electronic nose patterns of Korean red ginsengs were significantly different from those of Chinese red ginsengs. (iii) Some unidentified peaks were detected not in the headspace of Korean red ginsengs but in the headspace of Chinese red ginsengs when the headspace volatiles prepared by the SPME technique were analyzed by GC/ECD. (iv) Either the content ratios of K to Ca or Mn to Fe were significantly different between Korean red ginsengs and Chinese red ginsengs. (v) The reflectance ratios of NIRs wavenumbers such as $904\;cm^{-1}\;to\;1088\;cm^{-1}$ for Korean red ginsengs were significantly different from those for Chinese red ginsengs. (vi) The content ratios of ginsenoside-Rg to ginsenoside-Re of Korean red ginsengs were significantly higher than those of Chinese red ginsengs. These results indicate that the rhizome strength, the electronic nose pattern, the occurrence of ECD-sensitive headspace volatile components, the content ratios of K to Ca and Mn to Fe, the NIRs pattern and the content ratio of ginsenoside-Rg to -Re may be indicators for the discrimination of ginseng habitat.

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출류(朮類) 한약재의 외.내부형태와 이화학패턴 연구 (A Study on External.Internal Morphology and Pattern Analysis of Atractylodes Rhizomes)

  • 김정훈;이금산;최고야;황성연;김홍준;정승일;주영승
    • 대한본초학회지
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    • 제24권2호
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    • pp.77-85
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    • 2009
  • Objectives : To determine the discriminative criteria for Atractylodes rhizomes, the experiment of externalinternal characteristics and physicochemical pattern analysis were performed. Methods : External characteristics was observed using stereoscope. The sectioned materials which were covered with parffin were stained by Ju's method. Physicochemical patterns were analyzed using HPLC/DAD. Results : 1. External shape of original plant : Atractylodes maaocephala and A. japonica had relatively long petioles and 3-5 parted leaves. A. macrocephala had big purple flowers whereas A. japonica had relatively small white flowers and pinnate bracts. A. lancea had sessile leaves and white flowers, and the end parts of degenerated stamens were bent. 2. External shape of herbal medicine: A. macrocephala which was fist-shaped rhizome had pa-pillate processes and the cross section was light gray and sulcate. A. japonica and A. lancea were connected-beady or tubercular rhizomes, and the cross sections were both yellow-colored white. However, the cross section of A. japonica was fibrous, the width of cortex was narro-wer than that of stele, and radial shape in cortex was rare, whereas the width of cortex in A. lancea was similar to that of stele in size, and radial shape in cortex was obvious. 3. Internal shape of herbal medicine: A. macrocephala and A. lancea did not have lignified fascicles in cortex. However, the vascular bundles and vessels of A. macrocephala were wedge shaped and radial arrangement, and vascular bundles were densely populated in stele whereas those of A. lancea were repeatedly arranged and thinly extended to cortex. A. japonica had lignified fascicles in cortex and the width of vascular bundles was conspicuously thick with narrow intervals. 4. Physicochemical pattern analysis : A. macrocephala and A. lancea contained atractylenolide I and atractylenolide Ill whereas A. japonica contained atractylenolide I, atractylenolide Ill. diacetyl-atractylodiol, compound-4, compound-5. The three species of Atractylodes rhizomes showed different chromatogram patterns. Conclusions : The results could be used as discriminative criteria for Atractylodes rhizomes and as fundamental materials to researches of further pattern analysis and biological reaction.

콩나물중 살균제 carbendazim 잔류분의 정량 및 확인 (Determination and confirmation of the carbendazim residue in soybean sprout)

  • 김영국;박종태;홍석순
    • 농약과학회지
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    • 제2권3호
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    • pp.79-84
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    • 1998
  • 콩나물중 carbendazim 잔류분을 정량하고 확인할 수 있는 새로운 분석법을 확립하고자 tandem HPLC(UV & FL) 및 APcI를 source로 사용한 LC/MS를 이용하였다. 이를 위해 FL(fluorescence) 검출기를 UV(ultraviolet) 검출기와 나란히 연결하여 UV 검출기의 경우 280 nm 파장을 그리고 FL 검출기의 경우는 excitation파장과 emission파장을 각각 280 mn와 310 nm로 설정하였다. 분석결과 carbendazim의 검출한계는 $0.04{\mu}g/kg$이었다. 콩나물에 carbendazim을 0.5, 1.0 및 2.0 ppm 수준으로 첨가하여 회수율을 측정한 결과 그 평균값은 89.1%이었다. APcI source를 사용한 LC/MS 질량스펙트럼 방법은 콩나물중 carbendazim 잔류분을 최종 확인할 수 있었다. APcI LC/MS 방법은 전자충격에 의한 질량스펙트럼에 비해 훨씬 간단한 fragment를 형성할 뿐 만 아니라 carbendazim의 경우 m/z 133, m/z 159, m/z 191($M^{+}$)의 전형적인 fragment 이온을 형성하므로, 이 방법을 병행한다면 콩나물중 carbendazim 잔류분을 효과적으로 확인할 수 있을 것으로 사료되었다.

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가미청상보하탕(加味淸上補下湯) 열수추출물(熱水抽出物)과 알코올추출물(抽出物)의 지표물질(指標物質) 및 패턴의 비교분석(比較分析) (Comparison and Analysis of Several Main Components between Hot Water Extract and Alcohol Extract of Gamichungsangbohatang)

  • 민정기;정희재;이형구;정승기
    • 대한한방내과학회지
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    • 제27권1호
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    • pp.55-71
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    • 2006
  • Objectives : To find out more pharmacologically efficient way of extraction of herbal dicoction, Gamichungsangbohatang (GMCSBHT). Methods : Several main components of GMCSBHT was compared and analysed between hot water extract of GMCSBHT and Alcohol(70% ethanol) extract of GMCSBHT via HPLC method. Results : Hot water extract of GMCSBHT showed relatively more component content than ethanol extract of GMCSBHT. Also weighted mean of main components of hot water extraction of GMCSBHT was higher than that of alcohol extract of GMCSBHT. But from chromatographic pattern analysis of matching ratio and similarity ratio showed that these two forms of extracts might have different chemical composition, and 3D PDA plot of alcohol extract of GMCSBHT showed high peaks near UV $190{\sim}220nm$ which was invisible in hot water extract of GMCSBHT. Conclusion : Alcohol extract of GMCSBHT may have some special components which do not exist in hot water extract of GMCSBHT.

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